摘要
本研究采用SD大鼠乳鼠心肌细胞原代培养,复制心肌细胞缺氧复氧损伤模型,考察琥珀酸对心肌细胞缺氧复氧损伤的LDH漏出率的影响,并进一步采用流式细胞术及Western blot考察琥珀酸对心肌细胞凋亡,cleaved caspase-3及p-Akt的影响,探讨琥珀酸对新生大鼠原代心肌细胞缺氧复氧损伤的保护作用。研究结果发现：①琥珀酸31.25~500 mgL-1对原代心肌细胞活力无明显影响,琥珀酸400,200,100,50 mgL-1均可显著降低心肌细胞缺氧复氧损伤LDH漏出率（P〈0.01或P〈0.05）;②琥珀酸400,200 mgL-1可显著降低缺氧复氧损伤所致的心肌细胞凋亡百分数（P〈0.05）,抑制心肌细胞缺氧复氧所致的cleaved caspase-3 蛋白表达增加（P〈0.05）;③琥珀酸400 mgL-1可显著增加心肌细胞的p-Akt蛋白表达（P〈0.05）,而琥珀酸200 mgL-1对p-Akt蛋白表达无明显影响。因此,本研究认为琥珀酸可通过激活Akt的磷酸化而抑制心肌细胞缺氧复氧所致的坏死和凋亡。
To establish cardiomyocyte hypoxia/reoxygenation injury model by culturing primary cardiomyocytes from suckling SD rats, in order to study the effect of succinic acid on LDH leakage rate cardiomyocyte ischemia/reperfusion injury. Furthermore, flow cytometry and western blot were conducted to detect the effect of succinic acid on cardiomyocyte apoptosis, cleaved caspase-3 and p-Akt, and discuss the protective effect of succinic acid on primary cardiomyocyte hypoxia/reoxygenation injury of primary cardiomyocytes from neonatal SD rats. According to the findings of the study, succinic acid at the concentrations ranging between 31.25 mgL-1 and 500 mgL-1 had no significant effect on primary cardiomyocyte activity, and succinic acid at the concentrations of 400, 200, 100, 50 mgL-1 could notably reduce cardiomyocyte ischemia/reperfusion LDH leakage rate （P〈0.01 or P〈0.05, respectively）. Succinic acid at the concentrations of 400 mgL-1 and 200 mgL-1 could significantly reduce the percentage of cardiomyocyte apoptosis （P〈0.05）, and inhibit the protein expression of cleaved caspase-3 caused by cardiomyocyte ischemia/reperfusion （P〈0.05）. Succinic acid at the concentration of 400 mgL-1 could remarkably increase the protein expression of cardiomyocyte Akt （P〈0.05）, while succinic acid at the concentration of 200 mgL-1 had no obvious effect on the protein expression of cardiomyocyte Akt. Therefore, this study demonstrated that succinic acid could inhibit necrosis and apoptosis caused by cardiomyocyte hypoxia/reoxygenation by activating Akt phosphorylation.keywords：succinic acid
出处
《中国中药杂志》
CAS
CSCD
北大核心
2013年第21期3742-3746,共5页
China Journal of Chinese Materia Medica
基金
国家"重大新药创制"科技重大专项(2012ZX09301002-004-002)
国家自然科学基金项目(81073085
81001662)
关键词
琥珀酸
心肌细胞
缺氧复氧
坏死
凋亡
succinic acid
cardiomyocyte
hypoxia/reoxygenation
necrosis
apoptosis
